畜牧与兽医2024，Vol.56Issue(3)：69-76,8.
重组CRM197-4GnRH去势疫苗抗原分子的原核表达及免疫效果评价
Prokaryotic expression and immunological evaluation of recombinant CRM197-4GnRH castration vaccine molecules
摘要
Abstract
The CRM197-4GnRH antigen was expressed using an Escherichia coli expression system and vaccines were prepared with differ-ent adjuvants for immunization of rats to determine their immunocastration effects,aiming to provide insights for the development of a geneti-cally engineered GnRH-based vaccine with optimal immunogenicity,prolonged efficacy,and cost-effectiveness for immunocastration in ani-mal models.The CRM197-4GnRH gene was linked to the expression vector using Diphtheria toxin CRM197(Corynebacterium Diphtheriae An-tigen,CRM197-Conjugate)in series with four copies of GnRH(gonadotropin releasing hormone),and the optimal expression conditions of Escherichia coli were determined by optimization test.The expression of the target protein was identified by SDS-PAGE and Western blot.Nine vaccines utilized for testing purposes were prepared by emulsifying the fusion protein with different adjuvants,and were then immunized with GnRH antibody titers and testosterone concentration in the serum of rats.The results showed that the optimal expression conditions of Escherichia coli were determined as follows:induction time,5 h;LB liquid,medium;IPTG induction concentration,0.5 mmol/L.The tar-get fusion protein was successfully expressed through SDS-PAGE and Western blot analysis.The results of the animal experiments showed that the group immunized with the recombinant vaccine formulated with water-in-oil-in-water(W/O/W)adjuvant,the group immunized with the recombinant vaccine formulated with oil-in-water(O/W)adjuvant,and the group immunized with the GnRH+Th(Helper T cells)epitope(G1)formulated with oil-in-water(O/W)adjuvant,all exhibited a significant increase in GnRH antibody titer and a significant decrease in testosterone concentration,compared with the other groups(P＜0.05).The present results indicate that the above three vaccines exhibited significant immunocastration effects,laying the foundation for the development of GnRH immunocastration vaccines.关键词
CRM197/GnRH/大鼠/免疫去势Key words
CRM197/GnRH/rats/immune castration分类
农业科技引用本文复制引用
高华义,王敏,刘堃,张璐,王永芳,任东兴,郭钰洁,武毅,申雁冰,付旭彬..重组CRM197-4GnRH去势疫苗抗原分子的原核表达及免疫效果评价[J].畜牧与兽医,2024,56(3):69-76,8.基金项目
国家科学技术部资助项目(2014DFA31180) （2014DFA31180）
天津市科技支撑重点项目(20YFZCSN00120) （20YFZCSN00120）
